Graham R W, Atkinson T, Kilburn D G, Miller R C, Warren R A
Department of Microbiology, University of British Columbia, Vancouver, Canada.
Nucleic Acids Res. 1993 Oct 25;21(21):4923-8. doi: 10.1093/nar/21.21.4923.
A synthetic gene encoding the Schizophyllum commune xylanase XynA was constructed by a novel PCR-based procedure. Three long oligonucleotides were synthesized and used in combination with flanking PCR primers to generate a 607 base pair gene which contained 31 unique locations for restriction enzyme cleavage. The amino acid sequence was tailored for expression in Escherichia coli by using only those codons found in highly expressed E. coli genes. The availability of the gene will facilitate analysis of the structure and function of this and other beta-(1,4) xylanases.
通过一种基于聚合酶链式反应（PCR）的新方法构建了编码裂褶菌木聚糖酶XynA的合成基因。合成了三条长寡核苷酸，并与侧翼PCR引物结合使用，以生成一个607个碱基对的基因，该基因包含31个用于限制性内切酶切割的独特位点。通过仅使用在高表达的大肠杆菌基因中发现的那些密码子，对氨基酸序列进行了优化，以便在大肠杆菌中表达。该基因的获得将有助于分析这种和其他β-（1,4）木聚糖酶的结构和功能。
